Re: [Histonet] RE: Histonet digest, Vol 1 #49

Dear Rosemarie, This is the procedure from Sheehan and Hrapchak: 200gm Potassium dichromate 1 liter Distilled water 750ml Concentrated sulfuric acid Use heat resistant glassware, as heat will be genenated when adding sulfuric acid to mixutre. The solution is dark red brown, and may be continued to be reused for soak until it turns dark green. Do your stirring with a glass rod, and use all safety precautions due to the highly corrosive nature of the mixture. I hope this will be of some help to you. Sincerely, Paul Lockwood 9/16/03 12:57:42 PM, "Behan, Rosemarie G" wrote: >I am looking for a recipe for acid cleaning glassware to do silver stains, >can anyone help me? > >-----Original Message----- >From: histonet-request@lists.utsouthwestern.edu >[mailto:histonet-request@lists.utsouthwestern.edu] >Sent: Tuesday, September 16, 2003 1:00 PM >To: histonet@lists.utsouthwestern.edu >Subject: Histonet digest, Vol 1 #49 - 19 msgs > > >Send Histonet mailing list submissions to > histonet@lists.utsouthwestern.edu > >To subscribe or unsubscribe via the World Wide Web, visit > http://lists.utsouthwestern.edu/mailman/listinfo/histonet >or, via email, send a message with subject or body 'help' to > histonet-request@lists.utsouthwestern.edu > >You can reach the person managing the list at > histonet-admin@lists.utsouthwestern.edu > >When replying, please edit your Subject line so it is more specific >than "Re: Contents of Histonet digest..." > > >Today's Topics: > > 1. Re: Bad Plants/Yarrow (sara goldston) > 2. background problem (Jacqueline Miller) > 3. Re: Bouin's fixation/removal of picric acid (Sarah Jones) > 4. eye morphology advice (Margaret Gondo) > 5. ASCP BOR results (Lee & Peggy Wenk) > 6. wage and salary survey (Lee & Peggy Wenk) > 7. Colloidal Iron (lena spencer) > 8. Re: Bouin's fixation/removal of picric acid (John Kiernan) > 9. Re: Colloidal Iron (John Kiernan) > 10. RE: eye morphology advice (Kemlo Rogerson) > 11. Re: Methacarn/Carnoy's Storage (Barbara Osborn) > 12. IHC on Blood Smears (Jackie.O'Connor@abbott.com) > 13. Re: eye morphology advice (Fred Underwood) > 14. Re: eye morphology advice (Fred Underwood) > 15. Re: IHC on Blood Smears (Fred Underwood) > 16. immunohistochemistry and frozen sections (Vincenzo La Manna) > 17. Re: immunohistochemistry and frozen sections (Phillip Huff) > 18. Talented Histotechs wanted (Tommy Vedilago) > 19. Re: immunohistochemistry and frozen sections >(nick.kirk3@btopenworld.com) > >--__--__-- > >Message: 1 >Date: Mon, 15 Sep 2003 12:43:52 -0700 (PDT) >From: sara goldston >To: "Johnston, Kathy" , > "Histonet \(E-mail\)" >Subject: Re: [Histonet] Bad Plants/Yarrow > >--0-349928651-1063655032=:6512 >Content-Type: text/plain; charset=us-ascii > >I use to be a naturalist in high school and yarrrow is suppose to have >medicinal purposes (hopefully you and I are talking about the same plant). >If you crush either the leaves or blooms, cannot recall which, under you >nose you will smell something similar to Vick's vapor rub. Yarrow has the >same effect as Vick's by opening up the nasal passages. But more to your >question at hand. Yarrow is a wild bush so I would assume like any wild and >un-domesticated plant, it will grow quite freely if given the opportunity >to. If you are from anywhere east of the Mississippi, there has been an >unusual amount of rain not to mention cooler than normal temperatures. If >your plant was kept outside, it would have loved the weather this season and >flourished! You may have to break out the pruining shears. Sorry, I don't >have any other helpful advice. Try going to a nature reserve, greenhouse or >horticulturalist, and perhaps someone along those lines will have some more >concrete advise. >Sara > >"Johnston, Kathy" wrote: >While we are talking "Bad Plants" I would like everyone's opinion on >Yarrow. I bought 2 plants this spring and they have grown into to very >large although quite stunning plants. I can't remember it's exact name >right now, but each stem is a different pastel color. My only concern with >is is that is started out in a 2" pot, and right now I would need a 5 gallon >pail to put it in if I dug it out! > >I am understanding that is seems quite prolific. But is it easy to control. > >Your honest opinions please!!!! > >Thanks! > >Kathy > > > > >--------------------------------- >Do you Yahoo!? >Yahoo! SiteBuilder - Free, easy-to-use web site design software >--0-349928651-1063655032=:6512 >Content-Type: text/html; charset=us-ascii > >

I use to be a naturalist in high school and yarrrow is suppose to have >medicinal purposes (hopefully you and I are talking about the same >plant). If you crush either the leaves or blooms, cannot recall which, >under you nose you will smell something similar to Vick's vapor rub. >Yarrow has the same effect as Vick's by opening up the nasal passages. >But more to your question at hand. Yarrow is a wild bush so I would >assume like any wild and un-domesticated plant, it will grow quite freely if >given the opportunity to. If you are from anywhere east of the >Mississippi, there has been an unusual amount of rain not to mention cooler >than normal temperatures. If your plant was kept outside, it would >have loved the weather this season and flourished! You may have to >break out the pruining shears. Sorry, I don't have any other helpful >advice. Try going to a nature reserve, >greenhouse or horticulturalist, and perhaps so > meone > along those lines will have some more concrete advise.

Sara

"Johnston, Kathy" ><Kathy.Johnston@CLS.ab.ca> wrote:

>
While we are talking "Bad >Plants" I would like everyone's opinion on Yarrow. I bought 2 >plants this spring and they have grown into to very large although quite >stunning plants. I can't remember it's exact name right now, but each >stem is a different pastel color. My only concern with is is that is >started out in a 2" pot, and right now I would need a 5 gallon pail to put >it in if I dug it out!
>

>
I am understanding that is >seems quite prolific. But is it easy to control.
>

>
Your honest opinions >please!!!!
>

>
Thanks!
>

>
Kathy
>

>Do you Yahoo!?
>href="http://us.rd.yahoo.com/evt=10469/*http://sitebuilder.yahoo.com">Yahoo! >SiteBuilder - Free, easy-to-use web site design software >--0-349928651-1063655032=:6512-- > > >--__--__-- > >Message: 2 >Date: Mon, 15 Sep 2003 15:02:24 -0500 >From: "Jacqueline Miller" >To: >Subject: [Histonet] background problem > >Hi, > >We're having trouble with high background when using Rat IgG1 antibodies = >(4 =B5g/ml) on paraffin-embedded sections of human tissues, with or = >without retrieval procedures. Does anyone have any advice? > >Thank you, > >Jacqueline Miller >Research Asst. I >OB/GYN Dept. >University of Texas Southwestern Medical Center >Dallas, TX > > > > > >--__--__-- > >Message: 3 >Date: Mon, 15 Sep 2003 15:57:36 -0500 >From: "Sarah Jones" >To: >Cc: , >Subject: Re: [Histonet] Bouin's fixation/removal of picric acid > >I use it on wet tissue, it does not appear that the lithium needs to be >removed. Maybe I'm not understanding your question. Sarah >Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page >47. > > >>>> John Kiernan 09/14/03 11:54PM >>> >Sarah, do you use the saturated lithium >carbonate in 70% ethanol on blocks of >tissue or on hydrated paraffin sections >that are yellow? > >If it's for blocks, how do you remove >lithium carbonate from the decolorized >specimens? > John Kiernan >-- >------------------------- >John A. Kiernan >Department of Anatomy and Cell Biology >The University of Western Ontario >London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ >___________ >Sarah Jones wrote: >> >> I've always used saturated lithium carbonate in 70% ethanol to >remove >> the picric acid. I believe it would be faster and more complete >than >> 70% alone. >> >> Sarah Jones HT(ASCP) >> Dept. of Vet. Anatomy & Public Health >> Histology Lab >> Texas A&M University >> College Station, TX 77843-4458 >> phone: 979-845-3177 >> fax: 979-458-3499 >> >> >>> "peptolab" 09/13/03 08:32AM >>> >> A 3-5 mm testicular biopsy should be fixed in one to two hours. Be >sure >> to >> rinse out as much yellow picric acid as possible (you can use >running >> water >> or 70% alcohol ). The basophilia of chromatin in the stored block >will >> deteriorate after some time if there is picric acid residue. >> >> Jeff Silverman HT HTL QIHC (ASCP) >> Southside Hospital >> Bay Shore NY >> > > >--__--__-- > >Message: 4 >Date: Mon, 15 Sep 2003 16:28:30 -0500 >From: "Margaret Gondo" >Reply-To: >To: >Subject: [Histonet] eye morphology advice > >Hi All - > >I have a grad student who is interested in preserving the >morphology of a monkey eyeball once it is removed. Does anyone >have any suggestions? Everything we have tried always results in >the posterior part of the eyeball caving in. > > > >Thanks in advance, >Margaret > > > > > > >--__--__-- > >Message: 5 >From: "Lee & Peggy Wenk" >To: "Histonet" >Cc: >Date: Mon, 15 Sep 2003 19:10:59 -0400 >Subject: [Histonet] ASCP BOR results > >For those interested in the pass rates of the HT and HTL and SLS ASCP exams >Jan. - June 2003, please read on. (For those not interested, please press >the DELETE key now.) > >Scaled score of 400 is required to pass the exam. For HT and HTL, both the >written (MCQ = Multiple Choice Question) and the practical portions must be >passed. > >NAACLS = National Accrediting Agency for Clinical Laboratory Sciences, which >is the accrediting agency for lab training programs. > >Total Number non-NAACLS candidates = Total Number taking exam - Total Number >NAACLS students > >HISTOTECHNICIAN > >HT MCQ >Mean = 428 >Range of Scores = 100-761 >Total Number Taking Exam = 206 >Total Pass = 111 (54%) >Total Number NAACLS students = 24 >Total Pass NAACLS students = 18 (75%) >Total Number non-NAACLS candidates = 182 >Total Pass non-NAACLS candidates = 93 (51%) > >HT Practical >Mean = 446 >Range of Scores = 201-889 >Total Number Taking Exam = 217 >Total Pass = 144 (66%) >Total Number NAACLS students = 24 >Total Pass NAACLS students = 20 (83%) >Total Number non-NAACLS candidates = 193 >Total Pass non-NAACLS candidates = 20 (64%) > >HT COMBINED >Total Number Taking Exam = 349 >Total Pass = 141 (40%) >Total Number NAACLS students = 23 >Total Pass NAACLS students = 15 (65%) >Total Number non-NAACLS candidates = 326 >Total Pass non-NAACLS candidates = 126 (39%) > >HT exam first given in 1948. Certified to date = 18,556 > > >HISTOTECHNOLOGIST > >HTL MCQ >Mean = 451 >Range of Scores = 100-646 >Total Number Taking Exam = 43 >Total Pass = 32 (74%) >Total Number NAACLS students = 0 > >HTL PRACTICAL >Mean = 487 >Range of Scores = 244-700 >Total Number Taking Exam = 54 >Total Pass = 43 (80%) >Total Number NAACLS students = 0 > >HTL COMBINED >Total Number Taking Exam = 78 >Total Pass = 39 (50%) >Total Number NAACLS students = 0 > >HTL exam first given in 1980. Certified to date = 2,070 > > >SPECIALIST IN LABORATORY SAFETY >Mean = 422 >Range of Scores = 363-511 >Total Number Taking Exam = 7 >Total Pass = 3 (43%) > >SLS exam first given in 2000. Certified to date = 138 > >Peggy A. Wenk, HTL(ASCP)SLS >William Beaumont Hospital >Royal Oak, MI 48073 > > > >--__--__-- > >Message: 6 >From: "Lee & Peggy Wenk" >To: "Histonet" >Cc: >Date: Mon, 15 Sep 2003 19:19:56 -0400 >Subject: [Histonet] wage and salary survey > >FYI - > >The September 2003 issue of "Laboratory Medicine" from ASCP has Part I of >the latest Wage and Salary Survey. This survey is done every other year. >This article shows national averages of all lab disciplines, and then breaks >it down into regional areas. Please remember, your state, or even your part >of the state, may pay higher or lower than these averages. > >Things of interest that I found: > >- HT and HTL still have the highest vacancy rates of all laboratory >personnel. This is the same as the survey done 2 years ago. > >- HTL and MT now have the same salary ranges, on average nationwide. In >other words, on national average, HTL are now being paid the same rate as >MT. > >A few of the charts on the national averages can be found on the ASCP Board >of Registry website: >http://www.ascp.org/bor/center/center_research.asp > >Thought some people might like to find their issue, or borrow one from a >colleague (or maybe even join ASCP). > >Peggy A. Wenk, HTL(ASCP)SLS >William Beaumont Hospital >Royal Oak, MI 48073 > > > >--__--__-- > >Message: 7 >From: "lena spencer" >To: >Date: Mon, 15 Sep 2003 20:58:58 -0400 >Subject: [Histonet] Colloidal Iron > >Hi All: >One of my Pathologist's has requested a Hale's Colloidal Iron for renal cell >carcinoma chromophobes. I am familiar with colloidal iron for >mucosubstances, but he feel's that this is not the same stain. The article >stated that the diagnosis is made from the colloidal iron stain or from >electron microscopy . This Pathologist is requesting that I post on the >histonet for additional information or for someone out in histoland who is >performing this procedure. >Your is insight would be greatly appreciated. >Lena > > > > >--__--__-- > >Message: 8 >Date: Tue, 16 Sep 2003 00:24:20 -0400 >From: John Kiernan >Reply-To: jkiernan@uwo.ca >To: Sarah Jones >CC: histonet@pathology.swmed.edu >Subject: Re: [Histonet] Bouin's fixation/removal of picric acid > >I didn't phrase my question very clearly, and >should have explained the reason for asking. > >Previously I've encountered lithium carbonate >for removing picric from sections. It's the >traditional thing to do after Bouin fixation >when the tissue is still yellow in the wax >and after hydration of the sections. >You can easily see when all the yellow has >gone. > >With a block you can't see the middle, and >I was wondering how long it takes until no >more yellow comes out into the liquid. With >70% and higher alcohols alone it seems to take >for ever (at least with 5 mm and larger specimens; >I haven't used tiny biopsies after Bouin fixation). >It would be nice to have an idea how much more >quickly the picric acid is extracted when the >70% alc is saturated with Li2CO3. > >I asked about removing the Li2CO3 because it's >almost insoluble in alcohol (Merck Index). That >would be 95-100% alc. I guess you rinse the >specimens in more 70% after extracting the >picric acid, and that removes the Li2CO3 and >you don't get damage from crystals forming in >the tissue. >-- >------------------------- >John A. Kiernan >Department of Anatomy and Cell Biology >The University of Western Ontario >London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ >_________________________ >Sarah Jones wrote: >> >> I use it on wet tissue, it does not appear that the lithium needs to be >> removed. Maybe I'm not understanding your question. Sarah >> Ref. Theory and Practice of Histotechnology, Sheehan, Hrapchak, page >> 47. >> >> >>> John Kiernan 09/14/03 11:54PM >>> >> Sarah, do you use the saturated lithium >> carbonate in 70% ethanol on blocks of >> tissue or on hydrated paraffin sections >> that are yellow? >> >> If it's for blocks, how do you remove >> lithium carbonate from the decolorized >> specimens? >> John Kiernan >> -- >> ------------------------- >> John A. Kiernan >> Department of Anatomy and Cell Biology >> The University of Western Ontario >> London, Canada N6A 5C1 >> kiernan@uwo.ca >> http://publish.uwo.ca/~jkiernan/ >> ___________ >> Sarah Jones wrote: >> > >> > I've always used saturated lithium carbonate in 70% ethanol to >> remove >> > the picric acid. I believe it would be faster and more complete >> than >> > 70% alone. >> > >> > Sarah Jones HT(ASCP) >> > Dept. of Vet. Anatomy & Public Health >> > Histology Lab >> > Texas A&M University >> > College Station, TX 77843-4458 >> > phone: 979-845-3177 >> > fax: 979-458-3499 >> > >> > >>> "peptolab" 09/13/03 08:32AM >>> >> > A 3-5 mm testicular biopsy should be fixed in one to two hours. Be >> sure >> > to >> > rinse out as much yellow picric acid as possible (you can use >> running >> > water >> > or 70% alcohol ). The basophilia of chromatin in the stored block >> will >> > deteriorate after some time if there is picric acid residue. >> > >> > Jeff Silverman HT HTL QIHC (ASCP) >> > Southside Hospital >> > Bay Shore NY >> > > > >--__--__-- > >Message: 9 >Date: Tue, 16 Sep 2003 01:22:32 -0400 >From: John Kiernan >Reply-To: jkiernan@uwo.ca >To: lena spencer >CC: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] Colloidal Iron > >There's just the one Hale's (1946) method, also called >the dialysed iron technique because if you make >the original reagent yourself you have to dialyse >the ferric hydroxide sol. Detailed instructions are >given in Pearse's Histochemistry (any edition). >There have been several variants of the principal >reagent. In all the techniques the bound ferric >iron is made visible by a Prussian blue reaction >with potassium ferrocyanide. > >In Chapter 10 of the latest (2002) edition of >Bancroft & Gamble's "Theory and Practice ..." by >Barbara Totty, the method attributed to Hale (1946) >uses a more easily made colloidal iron sol that >does not need to be dialysed. A more detailed >reading of Pearse (3rd ed) shows that this is close >to the reagent of G.Muller (3 papers in Acta Histochem >vol 2, 1955-56, not seen by me; they'll be in >German). This colloidal iron sol quickly became more >popular than Hale's reagent not only because it was >easier to make but also because it was more specific >for acid mucosubstances. The not-dialysed colloidal >iron solution, unlike Hale's, does not stain nuclei >and there is no "background" cytoplasmic blue colour >in renal tubules. > >Both Pearse and Totty point out that colloidal iron >staining does the same job as alcian blue (pH 2.5) >but provides a darker blue product and also some >nonspecific staining. It's more sensitive and less >clean. > >"Hale's colloidal iron" (subject of the original >enquiry) is probably not an appropriate name for this >technique, which has been significantly improved >since 1946. Pearse seriously questioned its >histochemical >specificity despite recognizing the improvements made >by Muller and several others. > >Alcian blue is easier to use and can be incorporated >into many other rational mixed staining procedures >for mucosubstances. I know nothing about renal >carcinoma >chromophobes. If they can be stained with colloidal >iron >they must be basophilic, not "chromophobic." There are >very few genuinely chromophobic (unstainable) >cytoplasms. >-- >------------------------- >John A. Kiernan >Department of Anatomy and Cell Biology >The University of Western Ontario >London, Canada N6A 5C1 > kiernan@uwo.ca > http://publish.uwo.ca/~jkiernan/ >__________________________________ >lena spencer wrote: >> >> Hi All: >> One of my Pathologist's has requested a Hale's Colloidal Iron for renal >cell >> carcinoma chromophobes. I am familiar with colloidal iron for >> mucosubstances, but he feel's that this is not the same stain. The >article >> stated that the diagnosis is made from the colloidal iron stain or from >> electron microscopy . This Pathologist is requesting that I post on the >> histonet for additional information or for someone out in histoland who is >> performing this procedure. >> Your is insight would be greatly appreciated. >> Lena > > >--__--__-- > >Message: 10 >From: "Kemlo Rogerson" >To: , > >Date: Tue, 16 Sep 2003 07:33:44 +0100 >Subject: RE: [Histonet] eye morphology advice > >Nitrocellulose and celloidin technique. Page 91 of Histopathologic >Technic and Practical Histochemistry. RD Lillie and Harold M. >Fullmer.1976, 1965 McGrew Inc. 1234567890 KPKP 7832109876 > >48hr Form fixation >dehydrate with successive baths of 35, 50, 65 & 80% alc. 24 hours each. >Open eye >Return to 80% alc. 24 hours >2x 100% alc 24 hours >6 hour bath in equal vols 100% alc and ether >Infiltrate 5-7 days in 10% nitrocellulose in 100% alc. >Infiltrate 5-7 days in 20% nitrocellulose in 100% alc >Place under bell jar and when surface is solid, examine daily, but lower >portion soft flood with chloroform 16-24 hours. >Pour off chloroform and let dry. >Attach to block with 20% nitrocellulose >Let dry and immerse in chloroform for several hours. >24 hour baths of 3:1 chloroform: cedar wood oil, 1:1 of the same, then >pure cedar wood oil. >Cut sections and store in 80% alc. > >Dangerous, smelly, time consuming but extremely good fun; definitely a >non-PC technique that works; done it myself as a pup. > >Mr Kemlo Rogerson MSc DMS MIBiol CBiol FIBMS >Tel: 01270 877625 >Mob: 07830 196072=20 >Mobile E-Mail kemlorogerson@3mail.com =20 >FAX & Answer Phone 0871 242 8094 >E-mail Accounts:=A0=20 >=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0=A0 kemlo@tiscali.co.uk=A0or=20 > kemlo1@btinternet.com=A0 >Disclaimer: The information contained in this message and/or any >attachments(s) may be of a private and confidential nature, and is >intended solely for the attention of the addressee. If you have received >this message in error or feel you should not have been the intended >recipient, please return it and any attachments to the sender >immediately. All messages relating to this communication should then be >deleted from your system. Unauthorised usage, copying, disclosure or >alteration of this message and/or attachment(s) is strictly prohibited. >Barking, Havering and Redbridge Hospitals NHS Trust will not be held >responsible for any direct or indirect damages which may arise from >alteration of this message or any attachment(s), by a third party or >resulting from the transmission of a virus. > >=A0 >=A0 >=A0 > > >-----Original Message----- >From: histonet-admin@lists.utsouthwestern.edu >[mailto:histonet-admin@lists.utsouthwestern.edu] On Behalf Of Margaret >Gondo >Sent: 15 September 2003 22:29 >To: histonet@pathology.swmed.edu >Subject: [Histonet] eye morphology advice > >Hi All - > >I have a grad student who is interested in preserving the=20 >morphology of a monkey eyeball once it is removed. Does anyone=20 >have any suggestions? Everything we have tried always results in=20 >the posterior part of the eyeball caving in. =20 > > > >Thanks in advance, >Margaret > > >=20 > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >--__--__-- > >Message: 11 >Date: Tue, 16 Sep 2003 08:10:58 -0400 (EDT) >From: "Barbara Osborn" >To: "Histonet" >Subject: [Histonet] Re: Methacarn/Carnoy's Storage > > >Tamara, > >You can store methacarn or Carnoy's for up to a couple of years as long >as you cap it very, very tightly to avoid evaporation of chloroform and >alcohol. So you can store a large batch of fixative and only take out >what is needed each week. After use, however, it must be discarded. > > >--__--__-- > >Message: 12 >To: "Histonet" >From: Jackie.O'Connor@abbott.com >Date: Tue, 16 Sep 2003 07:30:32 -0500 >Subject: [Histonet] IHC on Blood Smears > >This is a multipart message in MIME format. >--=_alternative 0044E0DC86256DA3_= >Content-Type: text/plain; charset="us-ascii" > >A colleague of mine would like to stain murine blood smears for Bcl2. Does >anyone have any experience doing IHC on blood smears which have been fixed >in Methanol? > >Thanks > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotheraputics >847.938.4919 >Jackie.O'Connor@abbott.com >--=_alternative 0044E0DC86256DA3_= >Content-Type: text/html; charset="us-ascii" > > >
A colleague of mine would like to stain murine >blood smears for Bcl2. Does anyone have any experience doing IHC on >blood smears which have been fixed in Methanol? >
>
Thanks >
>
Jacqueline M. O'Connor HT(ASCP)
>Abbott Laboratories
>Global Pharmaceutical Research and Development
>Discovery Chemotheraputics
>847.938.4919
>Jackie.O'Connor@abbott.com >--=_alternative 0044E0DC86256DA3_=-- > > >--__--__-- > >Message: 13 >Date: Tue, 16 Sep 2003 10:14:27 -0400 >From: "Fred Underwood" >To: >Subject: Re: [Histonet] eye morphology advice > >I think this was discussed in the past. If my memory serves me right, >injecting the eye with a gelatin solution before processing will help >retain the shape. A search of the archives may be worth while. > >Fred > >>>> "Margaret Gondo" 09/15/03 05:28PM >>> >Hi All - > >I have a grad student who is interested in preserving the >morphology of a monkey eyeball once it is removed. Does anyone >have any suggestions? Everything we have tried always results in >the posterior part of the eyeball caving in. > > > >Thanks in advance, >Margaret > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >--__--__-- > >Message: 14 >Date: Tue, 16 Sep 2003 10:14:27 -0400 >From: "Fred Underwood" >To: >Subject: Re: [Histonet] eye morphology advice > >I think this was discussed in the past. If my memory serves me right, >injecting the eye with a gelatin solution before processing will help >retain the shape. A search of the archives may be worth while. > >Fred > >>>> "Margaret Gondo" 09/15/03 05:28PM >>> >Hi All - > >I have a grad student who is interested in preserving the >morphology of a monkey eyeball once it is removed. Does anyone >have any suggestions? Everything we have tried always results in >the posterior part of the eyeball caving in. > > > >Thanks in advance, >Margaret > > > > > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > >--__--__-- > >Message: 15 >Date: Tue, 16 Sep 2003 10:26:51 -0400 >From: "Fred Underwood" >To: >Subject: Re: [Histonet] IHC on Blood Smears > >If the smears are quite thick and bloody, you may want to use a more >dilute hydrogen peroxide solution (1% or so) for quenching. The >vigorous reaction can strip material from the slide. Prolonged buffer >wahes are a good idea as well. > >Fred > >>>> 09/16/03 08:30AM >>> >A colleague of mine would like to stain murine blood smears for Bcl2. >Does >anyone have any experience doing IHC on blood smears which have been >fixed >in Methanol? > >Thanks > >Jacqueline M. O'Connor HT(ASCP) >Abbott Laboratories >Global Pharmaceutical Research and Development >Discovery Chemotheraputics >847.938.4919 >Jackie.O'Connor@abbott.com > > >--__--__-- > >Message: 16 >Date: Tue, 16 Sep 2003 15:48:43 +0100 (BST) >From: "Vincenzo La Manna" >To: >Subject: [Histonet] immunohistochemistry and frozen sections > > > >Hi Histonetters, > >I've been struggling for 4 months on my 5 microns >frozen sections trying to find estrogen receptors. >I've tried both immunoperoxidase kit from Vector >laboratories either indirect immunofluorecsence (FITC >Goat anti mouse Secondary Ab from Sigma) but I've not >yet obtained good results. >My primary Ab is a mouse monoclonal to bovine >Estrogen receptor from Cymbus biotechnology. >Sections come from tissue that is supposed to be a >non-target tissue (mid laminar region from cow's >hooves)blocking with 2% goat normal serum (sigma) >whash in tbs buffer >All different methods for antigen retrieval have been >tested but sections look poor regarding histologic >quality and I still have very big problems with non >specific binding an staining (indirect >immunofluorescence). >It looks like frozen sections are too tender to >efford the whole processing. >Every suggestion or reference is welcome, >thank U > >La Manna Vincenzo >Department of Agriculture and Forestry, University of >Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >Telephone; 01224 274259 >Fax; 01224 273731 >e-mail v.lamanna@abdn.ac.uk > > > > > >--__--__-- > >Message: 17 >Date: Tue, 16 Sep 2003 09:29:40 -0600 (MDT) >From: Phillip Huff >To: "Vincenzo La Manna" >Cc: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] immunohistochemistry and frozen sections > >First, I would recommend that you run some Western Blots to ensure that >your Antigen of interest is being expressed within your tissues. Once your >Western blots are positive, you can definetly say that your immunoreaction >is what needs to be optimized. > >Second, have you applied your immunoreaction technique to a known positive >tissue source, that has also been sectioned at 5 um? It may be a good >confirmation that your tissue may need to be processed differently or cut >thicker. > >Finally, if your end goal is immunolocalization and the immunohistochem is >not working, you may want to look into in situ hybridization. In your >tissue samples, the receptor may be quite sensitive to processing and you >may be losing a great deal of your antigen. I know very little about your >receptor, but does it have a high turnover rate? Perhaps it has been >degraded over time when frozen. Has the reaction been done in freshly >harvested tissues? Perhaps localizing the mRNA in the tissues may provide >you with the results you are looking for. Before doing this though, I >would confirm that your tissues express the mRNA of interest by doing some >RT-PCR. This will both confirm that your tissues have your target mRNA, >and if you use an internal housekeeping primer in the PCR, you can >determine the approximate copy number of your mRNA to help you in >determining how sensitive your in situ reaction must be. > >Hope this helps you out, > >Phil > > > >> Hi Histonetters, >> >> I've been struggling for 4 months on my 5 microns >> frozen sections trying to find estrogen receptors. >> I've tried both immunoperoxidase kit from Vector >> laboratories either indirect immunofluorecsence (FITC >> Goat anti mouse Secondary Ab from Sigma) but I've not >> yet obtained good results. >> My primary Ab is a mouse monoclonal to bovine >> Estrogen receptor from Cymbus biotechnology. >> Sections come from tissue that is supposed to be a >> non-target tissue (mid laminar region from cow's >> hooves)blocking with 2% goat normal serum (sigma) >> whash in tbs buffer >> All different methods for antigen retrieval have been >> tested but sections look poor regarding histologic >> quality and I still have very big problems with non >> specific binding an staining (indirect >> immunofluorescence). >> It looks like frozen sections are too tender to >> efford the whole processing. >> Every suggestion or reference is welcome, >> thank U >> >> La Manna Vincenzo >> Department of Agriculture and Forestry, University of >> Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >> Telephone; 01224 274259 >> Fax; 01224 273731 >> e-mail v.lamanna@abdn.ac.uk >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet >> > > > >--__--__-- > >Message: 18 >Date: Tue, 16 Sep 2003 08:31:16 -0700 >From: "Tommy Vedilago" >To: "Histonet (E-mail)" >Subject: [Histonet] Talented Histotechs wanted > >Hello Histonetters, > > If you have ever wanted to work for a lab that is truly a family = >atmosphere where your skills and abilities are concretely appreciated, I = >have just the job for you. I have 3 positions for Histotechs in Dallas, = >TX with a progressive lab that has been in place for 36 years now. They = >are offering outstanding pay, benefits, and atmosphere. They are also = >offering relocation and hire bonus. If this holds interest for you or if = >you know of someone in the job market, please give me a call at = >866-SYSTEM1. >Tommy Vedilago >System 1 Search >(864) 627-0012 >(864) 627-0013 Fax > > >--__--__-- > >Message: 19 >Date: Tue, 16 Sep 2003 16:58:35 +0100 (BST) >From: nick.kirk3@btopenworld.com >To: histonet@lists.utsouthwestern.edu >Subject: Re: [Histonet] immunohistochemistry and frozen sections > >If it's not an inpolite question, why are you trying to demonstrate estrogen >receptors on frozen tissue? >Is this absolutely neccessary as estrogen receptors are very easily >demonstrated on paraffin processed tissue, which maintains better morphology >and gets almost guaranteed results. > >We use the DAKO 1D5 clone on FFPE tissue using Vector Unmasking fluid for >Antigen Retrieval and the DAKO ChemMate detection system and get excellent >results every time. > >Nick Kirk >Histopathology >Hinchingbrooke Hospital >Huntingdon >England > >> from: Vincenzo La Manna >> date: Tue, 16 Sep 2003 15:48:43 >> to: histonet@lists.utsouthwestern.edu >> subject: Re: [Histonet] immunohistochemistry and frozen sections >> >> >> >> Hi Histonetters, >> >> I've been struggling for 4 months on my 5 microns >> frozen sections trying to find estrogen receptors. >> I've tried both immunoperoxidase kit from Vector >> laboratories either indirect immunofluorecsence (FITC >> Goat anti mouse Secondary Ab from Sigma) but I've not >> yet obtained good results. >> My primary Ab is a mouse monoclonal to bovine >> Estrogen receptor from Cymbus biotechnology. >> Sections come from tissue that is supposed to be a >> non-target tissue (mid laminar region from cow's >> hooves)blocking with 2% goat normal serum (sigma) >> whash in tbs buffer >> All different methods for antigen retrieval have been >> tested but sections look poor regarding histologic >> quality and I still have very big problems with non >> specific binding an staining (indirect >> immunofluorescence). >> It looks like frozen sections are too tender to >> efford the whole processing. >> Every suggestion or reference is welcome, >> thank U >> >> La Manna Vincenzo >> Department of Agriculture and Forestry, University of >> Aberdeen, 581 King Street, AB24 5UA, Aberdeen , UK. >> Telephone; 01224 274259 >> Fax; 01224 273731 >> e-mail v.lamanna@abdn.ac.uk >> >> >> >> >> _______________________________________________ >> Histonet mailing list >> Histonet@lists.utsouthwestern.edu >> http://lists.utsouthwestern.edu/mailman/listinfo/histonet > > > > >--__--__-- > >_______________________________________________ >Histonet mailing list >Histonet@lists.utsouthwestern.edu >http://lists.utsouthwestern.edu/mailman/listinfo/histonet > >End of Histonet Digest > > >LEGAL NOTICE >Unless expressly stated otherwise, this message is confidential and may be privileged. 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Re: [Histonet] RE: Histonet digest, Vol 1 #49 - 19 msgs